Jas. Hongo et al., Antibody binding regions on human nerve growth factor identified by homolog- and alanine-scanning mutagenesis, HYBRIDOMA, 19(3), 2000, pp. 215-227
The binding specificities of a panel of mouse monoclonal antibodies (MAbs)
to human nerve growth factor (hNGF) were determined by epitope mapping usin
g chimeric and point mutants of NGF. Subsequently, the MAbs were used to pr
obe NGP structure-function relationships. Six MAbs, which recognize distinc
t or partially overlapping regions of hNGF, were evaluated for their abilit
y to block the binding of hNGF to the TrkA and p75 NGF receptors in various
irt vitro assays, which included blocking of TrkA autophosphorylation and
blocking of NGF-dependent survival of dorsal root ganglion sensory neurons.
Three MAbs (911,912,938) were potent blockers of all activities, Potent bl
ocking of p75 binding occurs only with MAb 909, which recognizes an NGF reg
ion identified by mutagenesis as important for NGF-p75 binding, These resul
ts are consistent with recently proposed models of binding regions involved
in NGF-TrkA and NGF-p75 interactions generated through mutagenic analysis
and structure determination of the NGF-TrkA complex. These studies provide
insight to the epitope specificities and potency of MAbs that would be usef
ul for physiological NGF blocking studies.